Leak Testing USP40 Chap 1207.2

14/3/2019 © 2019 USPC Official 8/1/17 - 11/30/17 General Chapters: <1207.
2> PACKAGE INTEGRITY LEAK TEST TECHNOLOGIES
〈1207.2〉 PACKAGE INTEGRITY LEAK TEST TECHNOLOGIES
1. INTRODUCTION
2. DETERMINISTIC LEAK TEST TECHNOLOGIES
2.1 Electrical Conductivity and Capacitance (High-Voltage Leak Detection)
2.2 Laser-Based Gas Headspace Analysis
2.3 Mass Extraction
2.4 Pressure Decay
2.5 Tracer Gas Detection, Vacuum Mode
2.6 Vacuum Decay
3. PROBABILISTIC LEAK TEST TECHNOLOGIES
3.1 Bubble Emission
3.2 Microbial Challenge, Immersion Exposure
3.3 Tracer Gas Detection, Sniffer Mode
3.4 Tracer Liquid
1. INTRODUCTION
The purpose of this chapter is to provide information guiding the selection and proper use of leak test technologies (also called methodologies,
approaches, or methods). The leak test technologies described in this chapter were selected on the basis of relevant research study data
published in peer-reviewed journals and/or precision and bias study data generated in support of recognized test method standards. When
referencing standard test methods (e.g., ASTM), the reader is advised to refer to the most recent versions. In some cases, the scope of
referenced standard test methods does not include the package types of the scope in Package Integrity Evaluation—Sterile Products 〈1207〉. In all
cases, methods and literature studies are cited to provide benchmark information useful for pharmaceutical package leak test method
development and validation.
The technologies described in this chapter are not prescriptive methods but represent testing concepts that may be applied when leak testing
sterile product–packages. Test technologies vary in terms of their potential detection limits, reliability, and applications; therefore, none are
universally appropriate for leak testing all product–packages. This chapter provides information to allow a thorough comparison of testing
approaches so that the most appropriate technology for a given situation can be identified.
After a methodology has been selected for use, the test equipment operation and performance is qualified. Test method parameters are
optimized during method development and confirmed during validation. Thus, a final leak test method is specific to a particular container–closure
or product–package system.
The leak test methods included are divided into two categories: deterministic and probabilistic. Deterministic leak test methods (Table 1) are
preferred over probabilistic methods when other key method selection criteria permit. Probabilistic leak test methods (Table 2) are best used
when the product–package system proves incompatible with deterministic methods, or when method outcome requirements demand a particular
probabilistic testing approach.
In this chapter's Table 1 and Table 2, the “leak size detection limit” provided for each methodology refers to leakage rates/leak sizes listed by
row in Package Integrity Testing in the Product Life Cycle—Test Method Selection and Validation 〈1207.1〉, Table 1. The reported leak detection
limits were chosen on the basis of literature sources, as well as commonly accepted experience. This information is intended to aid in the selection
of the test technology but should not be used as a definitive statement of test method performance for any specific leak test method applied to
any given product–package system. Instead, leak detection limit and range should be established during leak test method development and
validation for the respective product–package or container–closure system. For instance, an approach cited as capable of detecting row 6 leaks
may be validated by the user to detect leaks as small as those in row 3. Conversely, a method described as capable of detecting leaks in row 3
may be determined by the user to detect leaks only as small as row 5.
This battery of testing technologies and the information provided are intended to aid, not limit, the selection, development, validation, and use
of leak test methods. Unlisted methodologies shown by the user to meet the qualification and validation requirements for a satisfactory leak test
may be used. In addition, listed technologies may demonstrate expanded testing capabilities beyond those currently identified.
Table 1. Deterministic Leak Test Technologiesa

Deterministic Package Measurement Effect of Test Time
Leak Test Content Package Leak Detection Outcome and Method Order of
Technologies Requirements Requirements Limitb Data Analysis on Package Magnitude
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14/3/2019 © 2019 USPC Official 8/1/17 - 11/30/17 General Chapters: <1207.2> PACKAGE INTEGRITY LEAK TEST TECHNOLOGIES

Quantitative
measure of
electrical
current
passing
through the
test sample:
provides an
indirect
determination
of leak
presence and
leak location
as shown by a
Liquid (with no drop in test
combustion sample
risk) must be Row 3 electrical
more resistivity, with
electrically Varies with a resultant Nondestructive,
Electrical conductive product– increase in although
conductivity than package. Less package, voltage impact of test
and electrically instrument, test reading above exposure on
capacitance Product must conductive sample fixtures, a product
(high-voltage be present at than liquid and method predetermined stability is
leak detection) leak site. product. parameters. pass/fail limit. recommended Seconds
Quantitative
measure of
gas headspace
content of the
test sample by
laser-based
gas analysis,
for a product
requiring a
headspace low
in oxygen,
carbon
dioxide, or
water; vapor
concentration;
and/or low in
absolute
pressure.
Gas volume,
path length, Whole test
and content sample
must be Row 1 leakage rate is
compatible determined by
with Varies as a compiling
Laser-based instrument’s Allows function of time readings as a
gas headspace detection transmission of span between function of
analysis capability. near-IR light. analyses. time. Nondestructive Seconds
a
All methods apply to nonporous, rigid and flexible packages as per the scope of 〈1207〉.
b
The leak detection limit cited for each technology refers to Package Integrity Evaluation—Sterile Products 〈1207〉, Table 1 and is provided for
information only. This information is intended to assist in early methodology selection. The validated leak detection limit for a product–package
test method may deviate from these values.

Quantitative
measure of
mass flow rate
resulting from
test sample
headspace
escape or
liquid product
volatilization
within an
evacuated test
chamber
housing the
test sample.
Quantitative
pressure
readings early
in the test
cycle indicate
larger leak
presence.
Gas or liquid Whole test

must be sample
present at leak leakage rate is
site. Presence determined by
of liquid at leak Row 3 comparing the
site requires test sample
test pressures Varies with mass flow
below vapor product– results to
pressure. Rigid, or package, results using
flexible with instrument, test leak rate
Product must package fixtures/chamber, standards and
Mass not clog leak restraint and method positive Seconds to
extraction path. mechanism. parameters. controls. Nondestructive minutes
Quantitative
measure of
pressure drop
within a
pressurized
test sample.
Pressure drop
readings are a
measure of
gas escape
through leak
paths.
Whole test
Gas must be sample
present at leak Compatible leakage rate is
site. with pressure determined by
detection Row 3 comparing Nondestructive, Minutes to
Product mode. pressure decay unless the days,
(especially Varies with results to means used to depending
liquids or semi- Rigid, or product– results using access test on package
solids) must flexible with package, leak rate sample interior volume and
not cover package instrument, and standards and compromises required
potential leak restraint method positive test sample leak limit of
Pressure decay sites. mechanism. parameters. controls. barrier. detection
a
b

Quantitative
measure by
spectroscopic
analysis of
tracer gas leak
rate emitted
from a tracer-
flooded test
sample
positioned in
an evacuated
Able to tolerate test chamber.
high-vacuum
Tracer gas test conditions Whole test
must be added sample
to package. Rigid, or leakage rate is Nondestructive,
flexible with calculated by unless tracer
Tracer gas package Row 1 normalizing gas
must have restraint the measured introduction
access to mechanism Varies with tracer leak into the
package instrument rate by tracer package
Tracer gas surfaces being Limited tracer capability and concentration compromises
detection, tested for gas test sample in the test test sample Seconds to
vacuum mode leaks. permeability fixtures. sample. barrier. minutes
Quantitative
measure of
pressure rise
(vacuum
decay) within
an evacuated
test chamber
housing the
test sample;
vacuum decay
readings are a
measure of
headspace
escape from
the test
sample, or
liquid product
volatilization.
Gas or liquid Whole test

must be sample
present at leak leakage rate is
site. determined by
comparing
Presence of Row 3 vacuum decay
liquid at leak results for the
site requires Varies with test sample to
test pressures product– results of tests
below vapor Rigid, or package, performed
pressure. flexible with instrument, test using leak rate
Product must package sample chamber, standards and
not clog leak restraint and method positive Seconds to
Vacuum decay path. mechanism parameters. controls. Nondestructive minutes
a
b
Table 2. Probabilistic Leak Test Technologiesa

Probabilistic Package Leak Measurement Effect of Test Time
Leak Test Content Package Detection Outcome and Method Order of
a
b
The leak detection limit cited for each technology refers to Package Integrity Evaluation—Sterile Products 〈1207〉, Table 1 and is provided
for information only. This information is intended to assist in early methodology selection. The validated leak determination limit for a
product–package specific test method may deviate from these values.

Qualitative
measure by visual
inspection of
bubble emission
caused by escape
of test sample
headspace while
sample is
submerged and
exposed to
Row 4 differential
Gas must be pressure
present at leak Varies with conditions.
site. product– Alternatively,
package, sample surfaces
Product test sample may be exposed
(especially fixtures and to surfactant.
liquids or semi- positioning,
solids) must Rigid, or method Continuous bubble
not cover flexible with parameters, emission indicates
package package and analyst leak presence,
Bubble surfaces to be restraint technique location, and
emission leak tested. mechanism. and skill. relative size. Destructive Minutes
Qualitative
measure by visual
inspection of
microorganism
growth inside test
samples filled with
growth-supportive
media or product,
post immersion in
heavily
contaminated
challenge media
while exposed to
Row 4 differential
pressure
conditions,
followed by
Varies with incubation to
Growth- container– encourage
supportive Able to tolerate closure, test microbial growth.
media or pressure and sample
product. immersion fixtures and Growth in the test
challenge. positioning, sample indicates
Presence of challenge the presence of
liquid at the Rigid, or condition test sample leak
Microbial leak site flexible with severity, and site(s) capable of
challenge, required for package inherent allowing passive
immersion method restraint biological or active entry of
exposure reliability. mechanism. variability. microbes. Destructive Weeks
a
b

Row 2
Varies with
test sample, Quantitative
method measure by
parameters, spectroscopic
test sample analysis of tracer
fixtures, and gas near the outer
Tracer gas analyst surfaces of the
must be added technique tracer-flooded test
to package. and skill. sample, sampled Nondestructive,
Smaller leak using a sniffer unless tracer
Tracer gas Leak site detection probe. gas
must have accessible to may be introduction to
access to probe. possible Tracer presence the package
package under above a pass/fail interior
Tracer gas surfaces to be Limited tracer optimum limit indicates compromises
detection, tested for gas test leak presence and test sample Seconds to
sniffer mode leaks. permeability. conditions. location. barrier. minutes
Measure of tracer
in test sample
previously
submerged in
tracer-charged
liquid while
exposed to
differential
pressure
conditions.
Alternatively,
tracer-charged
test samples may
Row 4 be submerged in
tracer-free
Varies with collection fluid.
container–
closure, test Tracer migration
sample measurement
fixtures and may be
positioning, quantitative (by
challenge chemical analysis;
condition preferred
severity, and approach for small
Rigid, or tracer liquid leak detection) or
flexible with content. qualitative (by
package visual inspection).
restraint Smaller leak
mechanism. detection Tracer presence
may be indicates leak
Able to tolerate possible site(s) capable of
Contents must liquid under allowing tracer
be compatible immersion. optimal test passage. Tracer
with liquid conditions magnitude may
tracer. Compatible employing indicate relative
with liquid chemical leak size
Product must tracer analysis (assuming a
not clog leak detection tracer single-leak Minutes to
Tracer liquid path. mode. detection. pathway). Destructive hours
a
b
2. DETERMINISTIC LEAK TEST TECHNOLOGIES
2.1 Electrical Conductivity and Capacitance (High-Voltage Leak Detection)
2.1.1
The electrical conductivity and capacitance leak test (high-voltage leak detection, or HVLD) is an approach for detecting the presence, and
potentially the location, of a leak(s) in the wall of a nonporous, rigid or flexible package containing liquid or semi-liquid product. Test analysis is
based on quantitative electrical conductance measurements (1–5). HVLD leak tests are generally nondestructive to the package and to the
product, although an evaluation of HVLD exposure impact on product physicochemical stability is advised.
The test is performed by first positioning the test sample (containing liquid product) onto an electrically grounded instrument test fixture.
Alternatively, the test sample may be placed onto a transport system that will carry the test sample through an electrically grounded testing
zone. Upon test start, an electrode uniquely designed for the product–package type under test exposes all or part of the test sample to a high-
frequency, high-voltage, low-amperage current. The presence of a leak path in the proximity of an electrically conductive, liquid-formulation
product results in a drop in the electrical resistance of the test sample, as shown by a spike in current passing through the test sample above a
predetermined pass/fail limit established using negative controls.
2.1.2
Rigid or flexible packages of nonporous components containing liquid or semi-liquid product may be tested:
Package components must be relatively electrically nonconductive.
Product must be electrically conductive, relative to the package.
Product must not be flammable (i.e., not a combustion risk).
Product must be near or at the leak inspection location at the time of the leak test.
Solidified, electrically conductive product that blocks leak paths may be detected.
Metal caps used to seal stoppered vial or cartridge packages conduct current, improving the likelihood of finding leaks under the cap.
HVLD tests are rapid, requiring no more than several seconds for a full scan of the test sample, thus making them appropriate for off-line
testing, or as an on-line, 100% product inspection test method. This technology is useful for any product life cycle phase.
2.1.3
HVLD instrumentation comes equipped with tooling and/or a test sample transport system for proper test sample and probe/ground
positioning, an internal high-voltage transformer, electrode voltage and ground potentiometer adjustment capabilities, and a test result output
display. The design and materials of construction used for the electrode probe and electrical ground are product–package specific.
2.1.4
The following are test parameters for the electrical conductivity and capacitance leak test, also known as HVLD:
Conductivity of test sample product relative to test sample package: a greater difference will improve leak detection sensitivity
Test voltage set point: voltage is set high enough to ensure leak detection, but not so high that current will arc, falsely rejecting the test
sample
Test sensitivity set point (potentiometer or gain set point): sensitivity should be maximized to ensure leak detection without triggering a
false reject result
Package content proximity to potential leak paths: leak detection sensitivity is directly related to the proximity of product to the leak
path
Electrode probe position relative to potential leak paths: probe proximity to the leak is directly related to the test method sensitivity
Speed at which the electrode passes over the test sample surface: although the test is very rapid, test speeds too rapid may cause leaks
to be missed
Moisture presence on the package: test sample surface condensation can potentially trigger a false reject reading
2.2 Laser-Based Gas Headspace Analysis
2.2.1
Gas headspace analysis via laser-based techniques provides a quantitative, nondestructive measure of oxygen content, water vapor content,
and low internal pressure in the headspace of a nonporous, rigid or nonrigid package (6–8). Some instruments are capable of measuring
headspace carbon dioxide concentration as well.
The test is performed by first placing the test sample in a fixture designed for precise test sample positioning. Upon test start, frequency-
modulated spectroscopy is used to cause a near-infrared (IR) diode laser light to pass through the gas headspace region of the sealed test
sample. Light is absorbed as a function of gas concentration and pressure. The absorption information is processed using phase-sensitive
detection techniques; a mixer demodulates the signal. The output voltage, which is proportional to the absorption line shape, is digitally
converted and further analyzed by a microprocessor, yielding test sample signal results. Final test sample readings are automatically generated
based on a comparison of test sample signals to a calibration curve. This curve is pre-established by using control packages flooded with
traceable gas reference standards. Gas headspace analysis, as a function of time, provides a quantitative measure of the total leakage rate of
the test sample. Leakage rates are judged acceptable or unacceptable on the basis of predetermined limits, calculated to ensure proper gas
headspace content maintenance over the product life cycle.
2.2.2
Rigid or flexible packages made of nonporous components (transparent or semi-transparent material, either amber or colorless) that allow
transmission of near-IR diode laser light may be tested. Test samples require a minimum headspace volume and headspace path length. The
requirements vary on the basis of the gas moiety to be tested and may be specific to the instrument as well as to the construction and design of
the package materials.
Test samples that may be analyzed fall into these categories:
Products that require low-oxygen or low-carbon-dioxide headspace content
Products that require low water vapor content (e.g., lyophilized or powdered products)
Products that require low internal package pressure (e.g., lyophilized products)
Package integrity, or absence of leakage, is confirmed by replicating tests on a given test sample as a function of time. Longer time periods
between tests are needed to detect smaller leaks. Mathematical models appropriate to leak flow dynamics may be used to predict the time
required for detecting leaks of various sizes or rates.
Headspace analysis at a single time point provides the headspace content result, which may or may not be indicative of package integrity.
A test result not meeting specification could be due to package leakage, or could result from improper package filling or assembly
processes that caused the package headspace to be out of specification.
A test result that meets specification may confirm package integrity if enough time has elapsed since product–package preparation for
measurable leakage to have occurred, assuming that the initial preparation of the test sample met manufacturing standards.
Methods of laser-based gas analysis may be used during any phase of the product life cycle. Tests are rapid and are appropriate for off-line
testing using lab-scale equipment (typical measurement time, 2 s) or as an on-line, 100% product inspection method (typical measurement
time, 0.2 s).
2.2.3
Test instrumentation for laser-based gas headspace analysis is capable of accurate and reproducible near-IR diode laser light emission, light
detection, and signal analysis. Tooling specific to product–package test samples is used to properly position test samples, ensuring reproducible
laser-light transmission and detection. Standards with components identical to the packages under test, in terms of both the materials of
construction and the dimensions (at the point where light is to be transmitted), are required. These standards also need to contain headspace
content that is representative of the gas mixture under test (i.e., oxygen, carbon dioxide, water vapor, pressure).
2.2.4
The following are test parameters for laser-based gas headspace analysis:
Test sample position with respect to laser beam transmission and detection points: imprecise test sample handling, and/or dimensional
irregularity of the package, can increase measurement standard deviation
Test sample headspace volume: the volume must meet minimum requirements for test instrumentation
Test sample headspace absolute pressure: headspace absolute pressure will influence the detection limit and range for all test types
Test sample speed: increase in testing speed will increase standard deviation of measurement
Test sample temperature: temperature can influence moisture and pressure test results (e.g., lower temperatures are associated with
lower internal pressure of the test sample) (9)
Test sample outer surface moisture: presence of moisture may hamper test performance
Time allotted between the replicate tests: performing replicate tests over a period of time allows calculation of continuous package
leakage
2.3 Mass Extraction
2.3.1
The mass extraction test is a nondestructive, quantitative measurement approach for detecting leakage in nonporous, rigid or flexible
packages (10). Leakage of package headspace gases and/or leakage below the product fill level may be detected, given appropriately designed
equipment and test parameters.
The test is performed by first placing the test sample inside a test chamber that is pneumatically connected to a mass extraction leak test
system equipped with a vacuum generator package. The test chamber is uniquely designed to contain the test package, which is fitted with
appropriate tooling to limit movement or expansion of moveable or flexible components, respectively.
Upon test start, the chamber is quickly evacuated for a predetermined time to reach a predetermined vacuum level. A series of such
evacuation cycles may be performed, each intended to identify smaller leakage rates. After each cycle, the test system is isolated from the
vacuum source and measurements of absolute pressure, pressure decay rate, and/or gas mass flow rate are captured. Readings greater than
predetermined limits that were established using negative controls are indicative of container leakage, triggering test cycle abort.
For those test samples passing all previous larger leak vacuum cycles, a final vacuum is drawn. The test system is then isolated from the
vacuum source. With all flow from the test chamber directed through the mass flow sensor, the mass flow rate is measured. Mass flow above a
predetermined limit established using negative controls is indicative of container leakage.
2.3.2
Nonporous, rigid or flexible packages may be tested. Packages containing gas, liquid, and/or solid materials can be tested:
Flexible packages or packages with nonfixed components require tooling to restrict package expansion or movement, respectively, when
exposed to test vacuum conditions. Tooling minimizes the seal stress of flexible packages and maintains consistent package volume and
differential pressure conditions across the leak path.
Gas headspace must be at atmospheric pressure or at a pressure notably greater than test vacuum conditions.
Package surfaces below the product-fill level may be leak tested for those solid-formulation products that do not block leak-site gas flow
and for those liquid products that volatilize at test vacuum but do not solidify and so block leak paths.
Packages ranging in volume from a few milliliters to several liters may be tested.
Tests require anywhere from several seconds to a few minutes to perform. Longer test times are necessary for testing larger-volume
packages. Lengthening test cycles also allows for detection of smaller leaks.
Mass extraction leak tests are useful in any phase of the product life cycle. Tests may be performed in a laboratory setting or off-line in the
production environment. Longer laboratory or off-line test cycle times are generally capable of detecting smaller leaks. Higher speed on-line
tests are restricted to larger leak detection.
2.3.3
Mass extraction test instrumentation consists of a system of conduits and valves that pneumatically connect a test chamber with a test system
pressure sensor, micro-flow mass sensor, and vacuum generator package, including an external vacuum source. The instrument includes
appropriate timers, electronic controls, and monitors. A fixed-size orifice is included for periodic system performance verification. The test
chamber is uniquely designed to contain the test package, which is fitted with appropriate tooling to limit movement or expansion of moveable
or flexible components, respectively.
2.3.4
The following are test parameters for mass extraction:
Pressures
— Test system pressure reading after initial gross leak check. At evacuation stage, pressure is a function of test system volume,
time allotted for evacuation, and the vacuum source pressure level.
— Test system pressure reading after the secondary evacuation stage(s): pressure above a predetermined limit is due to test
package leakage. The pressure level above baseline is a function of leak size, available headspace volume, and/or volatile liquid
in the test sample.
— The final absolute pressure of the test cycle must be lower than the headspace pressure of the test package for detection of gas
headspace leaks, and/or lower than the volatilization pressure of liquid product formulation for leaks located below the liquid-
product fill level.
Mass flow
— Mass flow rate reading after secondary evacuation stage(s).
— The mass extracted from the test sample is monitored for larger leak detection after the first secondary evacuation stage; mass
extracted from the test sample is monitored for the smallest leak detection after the last evacuation stage.
— Background flow level (i.e., the baseline flow or noise level) is the flow rate for packages without leaks. Baseline flow is a function
of test package and system outgassing, test system volume, and the time allowed for evacuation.
— The mass flow rate at steady-state conditions, when extracted from the test chamber, is equal to test sample leakage into the test
chamber, assuming that outgassing from the sample and external leakage from the test system are insignificant. Leakage is
identified once the mass flow rate notably exceeds the rate of negative controls.
Times
— Time allotted for system evacuation for gross leak detection: enough time is allotted to draw off most of the test chamber gases,
without exhausting the headspace gases of the test package, or without drawing off the liquid contents from grossly leaking
packages.
— Time allotted for large-leak check through the mass flow sensor: a brief time is required for detection of large- and medium-sized
defects.
— Time allotted for system evacuation for small leak detection: enough time should be allowed to establish the desired vacuum
equilibrium of the test chamber. Insufficient time will not adequately draw off gases sorbed onto package surfaces or entrapped
between components.
— Time allotted for mass flow to stabilize: after the secondary evacuation stage(s), monitor the flow as the flow rate approaches
steady state. Enough time is allotted so that flow from the smallest allowed defect is statistically greater than baseline (no-leak)
flow.
— Time allotted for the final leak test by mass flow: enough time should be allotted so that the mass flow rate exceeds baseline
readings for negative controls.
2.4 Pressure Decay
2.4.1
The pressure decay test is a quantitative measurement approach for detecting leakage in nonporous, rigid or flexible packages. The test is
destructive if the introduction of pressurized gas creates a break in the package wall or seal. The test is nondestructive if the introduction of gas
into the test sample does not compromise the package barrier. Pressure decay testing is intended for integrity testing of the gas headspace
region of the test sample.
To perform the test, a dry air or inert gas pressure source is attached to the test sample that is fitted with an internal pressure monitoring
device. The test sample is pressurized to a predetermined pressure, after which the pressure source is isolated from the test sample. The decay
in pressure is monitored for a predetermined time. Pressure decay that exceeds a predetermined limit established using negative controls
indicates container leakage.
The referenced ASTM F2095 method (11) is intended for testing flexible packages (pouches and foil-sealed trays). Seals or surfaces being
tested cannot be in contact with product such as water, oils, or other liquids. ASTM method A describes testing packages without use of a
restraint mechanism. The method requires that the package reach a stable volume configuration (i.e., it stops stretching) to take a
measurement. ASTM method B requires that the test sample is kept between restraining plates during the test to limit the volume of the
pressurized package.
2.4.2
Nonporous, rigid or flexible packages may be tested:
Package surfaces that can be tested are those unobstructed by product (e.g., oils, water, or other liquids); small leaks below the liquid-fill
level would not be detected by this method.
exposed to test pressure conditions. Tooling minimizes the seal stress of the flexible package and maintains consistent package volume
and differential pressure conditions across the leak path.
Pressure decay can be used for testing packages anywhere from a few milliliters in volume to large, bulk-storage vessels.
Tests require anywhere from a few seconds to a several hours to perform. Longer test times are necessary for testing larger-volume
containers. Lengthening test cycles also allows for detection of smaller leaks.
Pressure decay tests are useful in any phase of the product life cycle.
Tests may be performed in a laboratory setting or off-line in the production environment. Laboratory or off-line test equipment that allows for
longer test times is generally capable of detecting smaller leaks. Higher-speed, on-line pressure decay equipment may be used to check for
defects in open packages before package filling and closure.
2.4.3
Pressure decay test instrumentation includes conduits to connect the test sample with test system pressure transducers (absolute, differential,
or a combination of both) and a pressure source (12). Instrumentation includes appropriate timers, electronic controls, and monitors. Greatest
test method sensitivity and reproducibility are achieved when the instrument is kept in a temperature-controlled environment; test samples
(especially larger-volume samples) are kept at a controlled, constant temperature during test; and dry pressurizing gas at constant temperature
is used. It is optional to use tooling uniquely designed to limit movement or expansion of moveable or flexible components, respectively, thereby
keeping test sample volume constant and limiting seal stress.
2.4.4
The following are test parameters for pressure decay:
Test sample internal pressure after pressurization:
— The initial pressure reached after sample pressurization is a function of the test system volume, time allotted for pressurization,
pressure source capacity, and temperature of the test sample headspace.
— Higher pressure creates the potential for more rapid and sensitive leak testing. However, the selection of maximum test pressure
should take into consideration personnel safety risks and potential damage to equipment and package.
Pressure decay baseline: the baseline pressure decay (i.e., noise level) is the pressure drop that occurs for packages without leaks.
— Baseline pressure drop is a function of test package volume, temperature conditions, and the length of time allowed for pressure
to rise.
— Baseline pressure drop requiring longer time periods is affected by gas sorption onto test package surfaces, gas moisture content
(dry gas should be used), and gas temperature. Techniques to limit baseline pressure drop include the use of dry gases, and
keeping the test container and the pressurized gas at a constant temperature.
Pressure decay due to test package leakage: the extent of pressure decay above baseline is a function of leak size, available headspace
volume in the test sample, the initial pressure inside the test sample, temperature control, and the time allotted for pressure to rise.
Times
— Time allotted for test sample pressurization: enough time is allotted to establish the desired pressure inside the test sample.
— Time allotted after pressurization for pressure decay: enough time should be allotted so that the pressure decay from the smallest
leaks can be detected (i.e., baseline decay is exceeded).
— A time lag may be incorporated before monitoring for pressure decay to allow for gas equilibrium within the container and test
system.
Temperatures
— Temperature of the pressurized gas can significantly affect test method sensitivity and reliability, especially when testing larger-
volume containers.
— An increase in gas temperature causes a rise in pressure, and conversely, a rise in gas pressure triggers an increase in gas
temperature. Therefore, upon initial test package pressurization, the gas temperature will spike, causing a further spike in
pressure. The subsequent drops in temperature and pressure during system equilibrium may be mistaken for leakage.
— Effects of temperature variation may be minimized by: 1) allowing the temperature in the pressurized system to come to
equilibrium before starting the pressure decay test, 2) minimizing temperature variation outside the test system, 3) minimizing
test duration, and/or 4) applying a mathematical correction to the pressure readings.
2.5 Tracer Gas Detection, Vacuum Mode
2.5.1
The leak detection method for tracer gas detects leakage from nonporous, rigid or flexible packages. The method can be destructive or
nondestructive, depending on the test approach used. The test requires the presence of tracer gas inside the test sample package. Helium is the
most commonly used tracer gas, and hydrogen is also used. The leakage rate of tracer gas is quantitatively measured using a spectrometric
analytical instrument specific for the tracer gas. Instruments are designed to check for tracer gas leaking out of a test package, either by means
of a test chamber that can be evacuated to draw gas out of test sample leaks (the vacuum mode) or by use of a vacuum wand (the sniffer
mode) for scanning the outer surfaces of the test package (see Tracer Gas Detection, Sniffer Mode). The vacuum-mode tracer gas test is used
more commonly than the sniffer mode for integrity testing sterile pharmaceutical product–packages. The vacuum mode is both quantitative and
deterministic and is used to capture and quantify leakage from an entire test package, or it can be used to test for leakage along a test package
surface or seal, given proper sample fixtures. Both vacuum and sniffer testing modes using helium as the tracer gas are described in ASTM
F2391 (13).
To perform the vacuum mode test, test samples that have been fully or partially flooded with tracer gas are placed inside an evacuation
chamber that is pneumatically connected to the tracer gas analysis instrument. Alternatively, the test sample may be fitted in such a manner
that only the surface or seal of interest is exposed to the instrument, allowing for targeted leak detection at one specific seal or surface. In some
cases, test samples that cannot withstand the high-vacuum test conditions may be tested with the use of tooling to restrict package expansion
or movement.
At test start, the instrument's vacuum pump evacuates the test chamber or fixture, drawing leaking tracer gas through the analyzer. The
absolute leak rate of the test sample is calculated by normalizing test results by the partial pressure of the tracer gas within the test sample at
the time of test. For accurate results, tracer gas concentration within the sample must be uniform and consistent at the time of test; also, there
should be minimal tracer gas permeation out of the test sample that can mask test sample leakage. Calibration tracer gas reference standards
can be used for understanding the relationship between true leak rates and measured leak rates under actual test conditions.
The vacuum-mode tracer gas leak test is a nondestructive test, unless tracer gas introduction into the test sample requires package wall
compromise (e.g., piercing), or if the presence of tracer gas is detrimental to the package contents.
2.5.2
Rigid or flexible packages made of nonporous components:
Flexible packages or packages with nonfixed components may require tooling to restrict package expansion or movement, respectively.
Tracer gas permeation through the package material must not be so great that the leakage rate of concern is masked.
A wide range of package sizes may be tested.
Leak paths must be clear of liquid or solid materials that could potentially block tracer gas flow.
Caution is advised when testing liquid-filled packages, because vapors or liquid drawn into the test system can seriously damage
instrumentation.
Detection capabilities range from large leaks to the smallest leaks.
Method capability is related to the size of the unobstructed leak path.
Large leaks in the smallest packages may be missed because of the rapid loss of tracer gas (e.g., during the evacuation phase of the
vacuum mode test).
Significant tracer gas permeation through the package itself can interfere with the test by swamping leakage rate.
The test is nondestructive if the tracer gas is introduced into the package at the time of package assembly or closure, but the inclusion of
tracer gas may prevent introduction of these packages into commercial or clinical markets. The test is destructive if the introduction of tracer
gas compromises assembled package integrity (e.g., package puncture). Following test sample preparation, the actual leak test generally takes
less than 1 min.
Tracer gas leak test methods may find application in any product life cycle phase. They are generally used in a laboratory environment. Tracer
gas methods can also be used in production as an off-line testing approach; they can be used on-line if tracer gas is introduced into the test
samples before final package closure.
2.5.3
Analytical instrumentation specific for tracer gas detection (e.g., mass spectrometer for helium detection) is required, typically equipped with
an internal leak rate standard for instrument calibration at time of use. Additional equipment needed includes external calibration reference
standards of tracer gas, to be used for comparisons of true leak rate versus measured leak rate under actual test conditions; a tracer gas source
with a means for introducing tracer gas into the test sample; a test chamber or fixture to pneumatically connect the test sample with the
instrument; tooling to restrict package expansion or movement, as appropriate; a means for accessing and analyzing the test sample headspace
for tracer gas partial pressure after execution of the vacuum-mode leak test; and ventilation to remove tracer gas from the test area, or a tracer
gas recapture system.
2.5.4
The following are test parameters for tracer gas detection, vacuum mode:
Tracer gas partial pressure within the test sample at the time of test:
— When added by soaking the intact package, the concentration of the tracer gas is dependent on the time allotted, the positive
pressure applied, and the tracer gas leak rate, plus the rate of permeation into the package
— When added before package closure, tracer gas concentration is dependent on the gas flooding rate and time, the tooling and/or
enclosure used to concentrate gas inside the package, and the efficiency of package closure for preventing escape of tracer gas
— When added after package assembly, tracer gas concentration is dependent on the tooling used to pierce, flush, and vent the
package; the gas flooding rate and time; and the sealant material applied to reseal the puncture site
Differential pressure applied to the test sample during the vacuum mode will drive tracer gas out of the test sample, increasing method
sensitivity
Time allotted to allow for leakage to reach steady state should not exceed tracer gas permeation lag time
2.6 Vacuum Decay
2.6.1
The vacuum decay test is a nondestructive, quantitative measurement approach for detecting leakage in nonporous, rigid or flexible packages.
Leakage in the package headspace gas region and/or below the product-fill level may be detected given appropriately designed test parameters
and if product properties allow (as detailed below).
To perform the test, the test sample is placed in a closely fitting evacuation test chamber pneumatically connected to the leak test system,
which is equipped with an external vacuum source. The test chamber is uniquely designed to contain the test package. Test samples with
moveable or flexible components require appropriate tooling to limit the movement or expansion of such components, respectively.
Upon test start, the test chamber plus test system dead space are evacuated for a predetermined period of time. The targeted vacuum level
chosen for the test is predetermined on the basis of the test sample type, size, and content. The vacuum source is then isolated from the test
system. After a short time has elapsed to allow for system equilibration, the rise in dead space pressure (i.e., vacuum decay) is monitored for a
predetermined length of time using absolute and/or differential pressure transducers. A pressure increase that exceeds a predetermined
pass/fail limit established using negative controls indicates container leakage. ASTM F2338 may be referenced (14).
2.6.2
Nonporous, rigid or flexible packages may be tested. Packages containing gas, liquid, and/or solid materials can be tested:
exposed to test vacuum conditions. Tooling minimizes flexible package seal stress and maintains consistent package volume and
differential pressure conditions across the leak path.
Product–package gas headspace must be at atmospheric pressure or at a pressure notably greater than test vacuum conditions.
Package surfaces below the product-fill level may be tested for leaks for those solid dosage formulation products that do not block leak-
site gas flow or for those liquid dosage form products that volatilize at test vacuum without solidifying and blocking leak paths.
Packages ranging in volume from a few milliliters to several liters may be tested.
Vacuum-decay leak tests are useful in all phases of the product life cycle. Tests require anywhere from a few seconds to a few minutes to
perform. Longer test times are necessary for testing larger-volume packages or for detection of the smallest leaks. Longer test times are more
appropriately performed in a laboratory setting or off-line in the production environment. Higher-speed on-line equipment is generally used for
detecting larger leaks.
2.6.3
Vacuum-decay leak test instrumentation consists of a system of conduits and valves that pneumatically connect a test chamber with the test
system pressure sensors and an external vacuum source. The instrument includes appropriate timers, electronic controls, and monitors. An
external gas flow meter allows for periodic system performance verification. The test chamber is uniquely designed to closely contain the test
package and may be fitted with tooling to limit movement or expansion of moveable or flexible package components, as appropriate.
2.6.4
The following are test parameters for vacuum decay:
Pressures
— Test chamber pressure after evacuation: the initial pressure reached during test chamber evacuation is a function of test system
volume, time allotted for evacuation, and vacuum pump capacity. The absolute pressure of the test system must be lower than
the headspace pressure of the test package for detection of gas headspace leaks, and/or lower than the volatilization pressure of
the liquid-product formulation for leaks located below the liquid-product fill level.
— Pressure rise baseline: the baseline pressure rise (i.e., noise level) is the pressure increase that occurs for packages without
leaks. Baseline pressure rise is a function of test package and system outgassing, test system volume, and the time allowed for
pressure rise.
— Pressure rise due to test package leakage: the extent of pressure rise above baseline is a function of leak size, test chamber
vacuum level at test start, available headspace volume or volatile liquid in the test sample, and the time allotted for pressure
rise.
Times
— Time allotted for system evacuation: enough time should be allowed for establishing the desired vacuum pressure level of the test
chamber, plus draw off gases sorbed onto package surfaces or entrapped between components. Excessive time will evacuate
headspace from largely leaking packages, risking that there will be insufficient headspace gas for leak detection. Times should
not be so great that leaking package headspace gases are exhausted or the liquid product floods and contaminates the test
system.
— Time allotted after evacuation for pressure rise (vacuum decay): enough time should be allotted so that the pressure rise from
the smallest leaks to be detected exceeds baseline. A time lag may be incorporated after evacuation and before vacuum decay
monitoring to allow for gas equilibrium within the container and test system.
3. PROBABILISTIC LEAK TEST TECHNOLOGIES
3.1 Bubble Emission
3.1.1
The bubble emission leak test is a destructive, qualitative measurement approach for detecting and locating leaks in nonporous, rigid or
flexible packages containing headspace gas.
The test is performed in one of two ways. The first is an internal pressurization method referenced in ASTM F2096 (15) in which a positive
pressure air source with pressure monitor is inserted into the test sample. The test sample is then submerged in water, and air pressure is
applied to a predetermined level, for a predetermined time period. The second approach is referenced in ASTM D3078 (16). The intact test
sample is submerged in water or other suitable submersion fluid contained in a vacuum chamber. Vacuum is established to a predetermined
level, for a predetermined time period.
With both approaches, leakage can be observed as a continuous stream of bubbles emitted from the leak site. Bubble diameter and emission
rate may provide some indication of relative leak size. An alternative to test sample submersion is coating the test sample with surfactant, in
which case any leakage is seen as foaming or bubbling at the leak site. The surface tension of the submersion fluid or surfactant allows for
smaller bubble formation, potentially improving test sensitivity. Use of submersion fluid with low gas solubility may also improve test sensitivity.
The bubble emission test is categorized as a probabilistic leak test method. Although this method relies on the predictable flow of gas through
leak paths, escaping gas can become entrapped within or between package components; false-leak outgassing events may occur; gas emitted
from small leaks may solubilize in the immersion fluid before bubble formation; and test sample set up may be inadequate to ensure sufficient
differential pressure conditions and appropriate bubble visibility. The use of negative and positive controls along with test samples provides
evidence of test method limit of detection.
3.1.2
Nonporous, rigid or flexible packages with gas headspace may be tested by bubble leak methods:
Packages must be able to tolerate wetting or submersion
Flexible packages or packages with nonfixed components generally require tooling to restrict package expansion or movement,
respectively, when exposed to vacuum conditions. Tooling minimizes the seal stress of the flexible package and maintains consistent
package volume and differential pressure conditions across the leak path. However, tooling may block leak paths or hinder bubble
emission visibility
Only leak sites that are present in the gas headspace region of the package can be detected
This test is generally used for testing smaller-volume packages that are less than a few liters in size
Bubble tests are applicable in any product life cycle phase. Bubble tests require several minutes or longer for test sample analysis and
subsequent cleaning and/or drying. Bubble tests are most commonly used in laboratory settings as part of a research investigation to verify leak
presence and location. They can also be used as an off-line production leak test. Bubble tests are also used for integrity testing of aerosol–
package products in a research or production setting. In this application, the test may be considered nondestructive to the product–package test
sample.
3.1.3
Bubble tests require a pressure or vacuum source (as appropriate) equipped with pressure monitors and controls; tooling to restrict expansion
of flexible packages or movement of nonfixed components; submersion fluid or surfactant to be applied to the package surface; submersion
vessel equipped for external vacuum or internal pressure test mode; and visual inspection aids such as lighting, magnification, and/or
background, as needed.
3.1.4
The following are test parameters for bubble emission:
Differential pressure: greater differential pressure evokes more rapid bubble emission. Differential pressure should not be so great that
package seals are compromised or that gas escapes through large leaks so rapidly that it may be confused with package surface
outgassing
Time allotted during differential pressure application: longer test times allow for smaller leak detection
Times allotted for inspection (pacing) and for inspection breaks to lessen operator fatigue
Package positioning during inspection
Package mode of restraint for moveable or flexible components
Submersion fluid (or surfactant) surface tension: lower surface tension lowers method sensitivity
Inspection environment parameters: lighting intensity and angle, degree of magnification, background color
3.2 Microbial Challenge, Immersion Exposure
3.2.1
The immersion exposure microbial challenge is a destructive, qualitative measurement approach for confirming leaks in nonporous, rigid or
flexible packages.
The test is performed by first filling test samples with sterile, growth-supporting media, followed by incubation and visual inspection of
samples to ensure sample sterility before microbial challenge. Samples are then immersed in a concentrated bacterial suspension for a
predetermined time. Samples can be exposed during immersion to a predetermined vacuum for a predetermined time, followed by release of
vacuum while the packages remain immersed at ambient pressure for a predetermined time. Samples are then incubated under growth-
promoting conditions, followed by examination of package contents for evidence of microbial growth by visual inspection or other appropriate
analytical means. Alternative approaches can include exposure of immersed test samples to positive pressure conditions, or to multiple cycles of
vacuum and/or pressure conditions. Test sample leakage is evidenced by visible growth of the challenge microorganism(s) inside test samples.
Immersion microbial challenge tests rely on the presence of a liquid carrier in the leak path that sweeps microorganisms into the package or
provides a means whereby microorganisms can actively migrate and/or grow into the test sample. The use of negative and positive controls
along with test samples provides evidence of test method limit of detection.
The microbial challenge by immersion test is categorized as a probabilistic leak test because of the multiple events that must occur
sequentially and/or simultaneously for leak detection to take place. All such events are difficult to predict or control, especially for detection of
smaller leaks. For example, the microorganisms must be physically present at the leak site. The necessary presence of liquid in the leak path,
and/or flowing through the leak path, is influenced by the package materials of construction, leak path tortuosity and topography, media surface
tension, and leak path blockage by product, extraneous debris, or air locks. Microorganisms must not be hindered from entering the package by
getting trapped in a tortuous leak path, and enough microorganisms must enter the package to allow for sufficient growth that can be detected
visually after test sample incubation.
3.2.2
Microbial challenge tests by immersion are most useful when an appropriate and validated physicochemical leak test method does not exist, or
when the test outcome demands direct evidence of the prevention of microbial entry.
Nonporous packages of rigid or flexible components may be tested by immersion microbial challenge methods:
Packages must be able to tolerate submersion.
Tooling minimizes stress on flexible package seals and maintains consistent differential pressure conditions across the package seal.
Immersion microbial challenge tests are performed in a laboratory environment, not as an on-line test of the finished product.
A test requires several days to prepare; the test samples are pre-incubated before microbial challenge to ensure initial package content
sterility. The challenge itself, plus post-challenge sample decontamination, may take several hours. Final incubation followed by sample
inspection may take 1–2 weeks. The immersion microbial challenge test is primarily used in product–package development and validation
studies.
3.2.3
The test chamber for conducting the immersion microbial challenge test is designed and equipped to maintain suspension uniformity and
appropriate temperature while exerting the required differential pressure condition. Fixtures for restraining and/or positioning packages during
immersion are also required. Small, motile microorganisms are preferred for the challenge; examples include Brevundimonas diminuta and
Serratia marcescens. The immersion challenge media should support challenge microorganism growth to the desired concentration. Soybean-
casein digest medium is one commonly used medium.
The media filled into test samples may match the immersion challenge media formulation. Alternatively, product that has been shown to
support microbial growth may be used. Media should allow sufficient growth of the particular challenge organism so that package contamination
can be detected in the positive controls. Verification of growth promotion in the immersion media and the test sample media should be
performed each time the immersion challenge test is conducted (see Sterility Tests 〈71〉, Growth Promotion Test of Aerobes, Anaerobes, and
Fungi). Use of an incubation chamber is required for test samples, both before and after exposure. A means for determining microbial growth
inside test packages is required.
3.2.4
The following are test parameters for microbial challenge, immersion exposure:
Microorganism concentration: in the immersion challenge media, microorganism concentration should meet appropriate predetermined
levels throughout the challenge test (a minimum concentration of 105 CFU/mL is commonly used).
Media fill volume: the fill volume of media in test samples should be sufficient to ensure a liquid path at each potential leak site.
Exposure of package seals and leak sites to the media may also be accomplished by test sample positioning during the immersion
challenge. It is also necessary to have sufficient volume and correct composition of the package headspace to encourage growth of the
selected challenge microorganism. Note that when the composition of the package headspace does not include oxygen, other test
conditions (e.g., anaerobic) may be applicable.
Vacuum/pressure conditions: exposing immersed test samples to differential pressure conditions is an important method parameter that
serves multiple purposes:
— Differential pressure helps eliminate trapped air and ensures the presence of liquid media between package components and at
leak sites.
— Differential pressure simulates the pressure changes incurred during air or land freight transport of the product. Absolute
pressure conditions that correspond to various altitudes anticipated during land and/or air freight transport are provided in ASTM
D6653/D6653M (17). Differential pressure conditions of the test may be modified on the basis of knowledge of the product–
package shipping environment.
— Differential pressure exposure can simulate conditions experienced by the product during some sterilization treatments.
Test times: longer exposure times improve the likelihood of microbial ingress into defective test samples and positive controls. However,
a possible decrease in the ability of media to support growth over time must be considered.
— Time allotted for immersion exposure during differential pressure exposure (vacuum and/or pressure)
— Time allotted for immersion exposure at ambient pressure conditions
Temperature during challenge: temperatures sufficient to support microbial growth are recommended. Temperature cycling can also be
used as a tactic to eliminate airlocks and promote the presence of liquid media at package seal sites.
Pre- and post-challenge test incubation temperature and times: the temperature selected should allow sufficient microbial growth.
Incubation times should be sufficient to ensure visualization of growth; these times are determined on the basis of positive controls and
samples from growth-promotion studies.
Parameters for detecting post-incubation microbial growth (e.g., lighting and background color for visual inspection, handling procedure,
and pacing).
3.3 Tracer Gas Detection, Sniffer Mode
3.3.1
The following information is specific to the tracer gas detection performed in the sniffer mode. (For additional information, see Tracer Gas
Detection, Vacuum Mode.)
Tracer gas detection using a sniffer attachment is used to detect leak presence and location in nonporous, rigid or flexible packages. This is a
nondestructive leak test, unless tracer gas introduction into the test sample requires package wall compromise (e.g., piercing) or if tracer gas
presence is detrimental to package contents. The sniffer mode test using helium as the tracer gas is described in ASTM F2391, Procedure A (13).
Briefly, test samples are flooded completely or partially with the tracer gas via one of several options. These options may include piercing a
closed test sample to introduce pressurized tracer gas (sealant is applied to close the puncture site); flooding the test sample before package
closure; or “soaking” a closed test sample by pressurizing with tracer gas (most applicable to larger leak detection). Test samples are checked
for leakage by scanning the outer package surfaces using a vacuum wand that is pneumatically connected to the tracer gas analytical test
instrument (e.g., a mass spectrometer for helium detection). Calibration reference standards of tracer gas can be used for understanding the
relationship between true leak rates and measured leak rates under actual test conditions. The use of negative and positive controls along with
the test samples provides evidence of test method limit of detection.
The sniffer mode of tracer gas leak testing is a probabilistic leak test method. This is because the presence of concentrated tracer gas near the
test sample surface is not a well-defined or predictable event, and the sniffer scanning procedure is prone to variability related to human
technique. The sniffer mode is generally chosen when the leak location is to be identified.
3.3.2
Rigid or flexible packages made of nonporous components may be tested:
Flexible packages or packages with nonfixed components may require tooling or manual manipulation to force tracer gas through leak
paths.
Tracer gas permeation through the package material must not be so great that the leakage rate of concern is masked.
A wide range of package sizes may be tested, ranging from small packages to large multi-liter vessels.
Leak paths must be clear of liquid or solid materials that could potentially block tracer gas flow. The leak size detection capability is related to
an unobstructed leak path.
The sniffer probe must not be allowed to draw liquid or hazardous vapors into the test system, as this would risk serious instrument damage.
Tracer gas leak test methods require time for the introduction of tracer gas into the test package, and up to several minutes to scan the
package. Large leaks in the smallest packages may be missed because of the rapid loss of tracer gas.
Methods of tracer gas leak tests in the sniffer mode are generally used in the laboratory environment for locating package leaks. Tracer-gas
sniffer mode tests are useful in any product life cycle phase.
3.3.3
Test equipment for tracer gas leak detection is described in Tracer Gas Detection, Vacuum Mode. However, a sniffer probe with an enclosure
and/or tooling for concentrating tracer gas passing through smaller leaks is used, rather than the test chamber or test fixtures described in
Tracer Gas Detection, Vacuum Mode.
3.3.4
The following are test parameters for tracer gas detection, sniffer mode:
Differential pressure applied to the test sample will drive tracer gas out of the test sample, increasing the method's sensitivity. For
flexible packages, differential pressure can be exerted by package compression.
Tracer gas partial pressure within the package at the time of the test is discussed, with instructions, in the Tracer Gas Detection, Vacuum
Mode.
Aspects of the sniffer mode vacuum wand, such as sweeping speed and distance from the package surface, as well as the tooling or
enclosure used for concentrating leaking gas.
3.4 Tracer Liquid
3.4.1
The tracer liquid test method is a destructive approach for detecting and potentially locating leaks in nonporous, rigid or flexible packages.
Tracer liquid tests provide an indication of leak presence and may provide a measure of relative leak size. Tracer liquid tests using liquid
submersion work by the diffusive flow of the tracer element through a liquid-filled leak path and/or the effusive flow of tracer solution through
the leak path.
The liquid submersion test uses one of two basic approaches. In the first approach, test samples are submerged in a tracer-element solution
formulation contained in an evacuation chamber. Examples of tracer elements include dyes, radionuclides, or metallic ions. In the second
approach, test samples containing tracer formulation are submerged in tracer-free liquid contained in an evacuation chamber. For both
approaches, test samples may be fitted with tooling to ensure proper positioning and to restrict flexible or moveable components. The
submerged test samples are subjected to vacuum at a predetermined pressure level for a predetermined time. After vacuum release, test
samples remain submerged for a predetermined time. Additional test options include the use of positive pressure exposure or multiple cycles of
differential pressure conditions to encourage effusive flow of the tracer element through the leak path.
In the first approach, after the challenge is complete, test sample outer surfaces are cleaned, and the contents are checked for evidence of
tracer ingress (18). In the second approach, after the challenge is complete, the immersion fluid is checked for evidence of tracer liquid egress
out of the test sample. In both cases, measurement of tracer liquid migration may be performed in a quantitative manner by using chemical
analysis techniques (preferred for small leak detection). Alternatively, the presence of leakage may be determined qualitatively by visual
inspection if the tracer element can be discerned visually. The use of negative and positive controls along with test samples provides evidence of
test method limit of detection.
Liquid tracer tests are categorized as probabilistic methods. Successful liquid tracer detection relies on a combination of tracer solution
wicking, tracer solution effusion, and tracer element diffusion through a liquid-filled leak path and are events that are difficult to predict or
control, especially for detection of smaller leaks. These events are influenced by numerous factors, including the package materials of
construction, leak path tortuosity and topography, tracer liquid surface tension, and leak path blockage by product, extraneous debris, and air
locks.
3.4.2
Rigid or flexible packages of nonporous components may be tested using tracer liquid submersion methods:
Packages must be able to tolerate wetting or submersion.
— Testing by submerging a test sample in tracer liquid is used when the test sample allows for visual examination of the tracer (e.g.,
dye) ingress, or when the tracer element is to be contained within the test sample after testing.
— Testing by submerging tracer-filled test samples in tracer-free liquid may be used when the sample interior cannot be visually
examined or when the tracer element is best captured for analysis outside the test sample.
Tests may require up to 1 h or longer for test sample exposure, cleaning, and inspection or analysis. Tracer liquid tests are primarily used for
laboratory testing or off-line product sample testing. Tracer liquid submersion tests can be used in any product life cycle phase.
3.4.3
A test vessel equipped for challenge conditions of vacuum and/or positive pressure is required and is pneumatically connected to a pressure
and/or vacuum source, as appropriate, and equipped with pressure monitors and controls (19). The tracer liquid formulation should be
physicochemically compatible with the test sample components and the tracer-free solution to be filled into the test samples or used as the
immersion bath, to ensure optimal tracer solution functionality. Formulation considerations include tracer element type and concentration;
surfactant use, type, and concentration; and solvent system. Examples of tracer liquid incompatibilities apparent upon contact with the test
product include tracer dye fading, tracer element precipitation, and tracer element sorption onto package components. Tooling is often needed to
restrict flexible package expansion or nonfixed component movement upon differential pressure exposure. Tooling minimizes stress on flexible
package seals and ensures consistent differential pressure conditions across the leak path.
Tracer detection requires the use of either analytical detection instrumentation [e.g., UV-Vis spectrophotometry (20), phase induction
spectrophotometry, or other] or visual inspection aids. Analytical detection offers the advantage of minimizing the error that is inherent in visual
discernment of low dye concentrations. Optimal visual inspection requires the use of controlled inspection conditions, such as background color,
lighting, pacing, fatigue breaks, and negative controls for comparison.
3.4.4
The following are test parameters for tracer liquid:
Differential pressure conditions used (vacuum and/or pressure): greater pressure differentials encourages tracer liquid passage.
Submersion times during and after differential pressure application: longer times allow for greater tracer liquid passage through leak
paths.
Holding time between tracer liquid challenge and final inspection: some tracer liquids visibly fade or are sorbed onto package surfaces
over time.
Tracer liquid surface tension: lower surface tension allows for smaller leak detection.
Tracer detection parameters
— Analytical detection
Test sample content extraction procedure
Method-specific performance parameters
Visual inspection
Inspection environment parameters: lighting intensity and wavelength, background color, viewing angle, and test sample
visibility
Time allowed for inspection (pacing) and breaks to lessen operator fatigue
Comparison with negative controls
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General Chapter Radhakrishna S Tirumalai, Ph.D. (GCM2015) General Chapters-Microbiology 2015

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Previously Appeared In:
Pharmacopeial Forum: Volume No. 40(5)

Leak Testing USP40 Chap 1207.2

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14/3/2019 © 2019 USPC Official 8/1/17 - 11/30/17 General Chapters: <1207.

2> PACKAGE INTEGRITY LEAK TEST TECHNOLOGIES

〈1207.2〉 PACKAGE INTEGRITY LEAK TEST TECHNOLOGIES

2. DETERMINISTIC LEAK TEST TECHNOLOGIES

2.1 Electrical Conductivity and Capacitance (High-Voltage Leak Detection)

2.2 Laser-Based Gas Headspace Analysis

2.3 Mass Extraction

2.4 Pressure Decay

2.5 Tracer Gas Detection, Vacuum Mode

2.6 Vacuum Decay

3. PROBABILISTIC LEAK TEST TECHNOLOGIES

3.1 Bubble Emission

3.2 Microbial Challenge, Immersion Exposure

3.3 Tracer Gas Detection, Sniffer Mode

3.4 Tracer Liquid

Table 1. Deterministic Leak Test Technologiesa

Deterministic Package Measurement Effect of Test Time

Deterministic Package Measurement Effect of Test Time

Gas or liquid Whole test

Deterministic Package Measurement Effect of Test Time

Gas or liquid Whole test

Table 2. Probabilistic Leak Test Technologiesa

Probabilistic Package Leak Measurement Effect of Test Time

Probabilistic Package Leak Measurement Effect of Test Time

2. DETERMINISTIC LEAK TEST TECHNOLOGIES

2.1 Electrical Conductivity and Capacitance (High-Voltage Leak Detection)

2.2 Laser-Based Gas Headspace Analysis

2.3 Mass Extraction

2.4 Pressure Decay

2.5 Tracer Gas Detection, Vacuum Mode

2.6 Vacuum Decay

3. PROBABILISTIC LEAK TEST TECHNOLOGIES

3.1 Bubble Emission

3.3 Tracer Gas Detection, Sniffer Mode

3.4 Tracer Liquid

General Chapter Radhakrishna S Tirumalai, Ph.D. (GCM2015) General Chapters-Microbiology 2015

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